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目的:研究褪黑素对不同月龄小鼠免疫反应的影响及其作用机制。方法:淋巴细胞增殖采用MTT法;IL-2活性检测 采用激活小鼠脾淋巴细胞增殖法;cAMP测定用竞争蛋白结合法;甲硫氨酸脑啡肽的测定采用放免法。结果:6月龄和11月龄BALB/c小鼠胸腺淋巴细胞增殖能力和IL-2的产生均降低,褪黑素(5mg/kg或30mg/kg,ig×7d)对上述指标有不同程度的改善作用,在体外,11月龄小鼠胸腺淋巴细胞增殖反应明显降低,而其产生的cAMP则明显升高,褪黑素(0.1nmol/L和1μmol/L)对此有反向调节作用,弗司可林(10μmol/L)(腺苷酸环化酶选择性激活剂)能够增强2月龄和11月龄BALB/c小鼠胸腺淋巴细胞内的cAMP水平,褪黑素可部分拮抗此作用,褪黑素的这一作用可被百日咳毒素(1mg/L)完全取消,同时发现褪黑素(1μmol/L和0.1nmol/L)能够明显提高2月龄和11月龄BALB/c小鼠胸腺淋巴细胞的甲硫氨酸脑啡肽含量,这一作用能被硝苯地平(1μmol/L)所阻断,提示褪黑素促进淋巴细胞甲硫氨酸脑啡肽释放可能由Ca~(2+)通道介导。结论:褪黑素对不同月龄小鼠具有免疫调节作用;G蛋白偶联的AC-cAMP信号转导通路和淋巴细胞甲硫氨酸脑啡肽的释放可能是褪黑素发挥免疫调节作用的重要机制之一。
Objective: To study the effect and mechanism of melatonin on immune response in mice of different age. Methods: The proliferation of lymphocytes was detected by MTT method. The activity of IL-2 was detected by activated spleen lymphocyte proliferation method. The cAMP was measured by competitive protein binding method. The methionine enkephalin was determined by radioimmunoassay. Results: The proliferation of thymus lymphocytes and the production of IL-2 in 6-month-old and 11-month-old BALB / c mice were all decreased. The levels of melatonin (5mg / kg or 30mg / kg and ig × 7d) , The thymus lymphocyte proliferative response was significantly decreased in 11-month-old mice and the cAMP production was significantly increased in vitro. Melatonin (0.1nmol / L and 1μmol / L) had a reverse regulatory effect on this , And forskolin (10 μmol / L) (adenylate cyclase-selective activator) increased cAMP levels in thymocytes of 2-and 11-month-old BALB / c mice. The effect of melatonin was completely abolished by pertussis toxin (1 mg / L), and melatonin (1 μmol / L and 0.1 nmol / L) significantly increased BALB / c at 2 and 11 months Methionine enkephalin content in mouse thymus lymphocytes was blocked by nifedipine (1μmol / L), suggesting that melatonin may promote the release of methionine enkephalin from lymphocytes ~ (2+) channel mediated. CONCLUSION: Melatonin has immunomodulatory effects on mice of different ages. The release of G-protein-coupled AC-cAMP signal transduction pathway and lymphocyte methionine enkephalin may play an immunomodulatory role in melatonin One of the important mechanisms.